The conjugation of diphtheria toxin T domain to poly(ethylenimine) based vectors for enhanced endosomal escape during gene transfection.
The endosomal escape is a well-known serious obstacle for non-viral gene delivery. This is because of an acidic and enzymatic degradation of the contents of the endosome/lysosome. Therefore, the internalized gene needs to be efficient released into the cytosol to obtain the efficiently transfection efficiency. On the other hand, the diphtheria toxin T domain fuses with endosome membrane by pH decrease, then enhances the endosomal escape of the diphtheria toxin C fragment. In this study, we constructed diphtheria toxin T domain-conjugated poly(ethylenimine)s (PEI) polyplex for enhancing the endosomal escape of exogenous gene. The conjugation of diphtheria toxin T domain with PEI/pDNA polyplex leads to the significant enhancement of transfection efficiency when compared with plain PEI/pDNA polyplex. The pH-responsive increase in hydrophobicity of the diphtheria toxin T domain might not only trigger the perturbation of the endocytic vesicle membrane but might also increase the membrane permeability.
Kakimoto
Shinji
S
Department of Applied Chemistry and Bioengineering, Graduate School of Engineering, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan.
Hamada
Tsutomu
T
Komatsu
Yuuki
Y
Takagi
Masahiro
M
Tanabe
Toshizumi
T
Azuma
Hideki
H
Shinkai
Seiji
S
Nagasaki
Takeshi
T
D000818:Animals; D001692:Biological Transport; D019921:Biotinylation; D019556:COS Cells; D002463:Cell Membrane Permeability; D002522:Chlorocebus aethiops; D004167:Diphtheria Toxin / Q000737:chemistry* / Q000378:metabolism*; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D042541:Intracellular Space / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D011094:Polyethyleneimine / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D019809:Streptavidin / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D004167:Diphtheria Toxin; D008081:Liposomes; D011094:Polyethyleneimine; D019809:Streptavidin